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Key Specifications Table
| Key Applications | Species | Source |
|---|---|---|
| CULT | Human | Recombinant human DNA expressed in baculovirus infected Sf9 cells |
| Description | |
|---|---|
| Catalogue Number | 01-185 |
| Brand Family | Upstate |
| Trade Name |
|
| Description | VEGF Protein, Human recombinant |
| Product Information | |
|---|---|
| Formulation | |
| Presentation | 10 µg of VEGF lyophilized protein from buffer containing 25mM HEPES and 150mM NaCl, pH 7.0. Carrier free. |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | The human recombinant VEGF Protein is available in a 10 µg format. |
| Key Applications |
|
| Application Notes | VEGF Proliferation Assay 1. Prior to use in this assay, maintain HUVE cells in F-12K medium supplemented with 10% (v/v) FBS, 1% penicillin-streptomycin-glutamine, 0.1 mg/ml heparin and 0.05 mg/ml endothelial cell growth supplement (ECGS). 2. For assay of VEGF, subculture HUVE cells into a 96-well plate at a density of 4,000 cells per well in the growth medium over night. 3. Add VEGF to a final concentration range of 0.2, 0.4, 0.8, 1.6, 3.2, 6.2, 12.5, 25 and 50ng/ml. Add VEGF diluent to one set of wells as the negative control. 4. Four days later, add 20ul of Promega Substrate Cell Titer 96 Aqueous One Solution Reagent to each well. 5. Incubate at 37C for 2 hrs and read at OD 490nm. |
| Biological Information | |
|---|---|
| Source | Recombinant human DNA expressed in baculovirus infected Sf9 cells |
| Entrez Gene Number |
|
| Entrez Gene Summary | This gene is a member of the PDGF/VEGF growth factor family and encodes a protein that is often found as a disulfide linked homodimer. This protein is a glycosylated mitogen that specifically acts on endothelial cells and has various effects, including mediating increased vascular permeability, inducing angiogenesis, vasculogenesis and endothelial cell growth, promoting cell migration, and inhibiting apoptosis. Elevated levels of this protein is linked to POEMS syndrome, also known as Crow-Fukase syndrome. Mutations in this gene have been associated with proliferative and nonproliferative diabetic retinopathy. Alternatively spliced transcript variants, encoding either freely secreted or cell-associated isoforms, have been characterized. There is also evidence for the use of non-AUG (CUG) translation initiation sites upstream of, and in-frame with the first AUG, leading to additional isoforms. |
| Gene Symbol |
|
| Purification Method | Sterilized through a 0.2 micron membrane filter |
| UniProt Number |
|
| UniProt Summary | FUNCTION: SwissProt: P15692 # Growth factor active in angiogenesis, vasculogenesis and endothelial cell growth. Induces endothelial cell proliferation, promotes cell migration, inhibits apoptosis, and induces permeabilization of blood vessels. Binds to the VEGFR1/Flt-1 and VEGFR2/Kdr receptors, heparan sulfate and heparin. Neuropilin-1 binds isoforms VEGF-165 and VEGF-145. SIZE: 232 amino acids; 27042 Da SUBUNIT: Homodimer; disulfide-linked. Also found as heterodimer with PlGF (By similarity). SUBCELLULAR LOCATION: Secreted. Note=VEGF121 is acidic and freely secreted. VEGF165 is more basic, has heparin-binding properties and, although a signicant proportion remains cell-associated, most is freely secreted. VEGF189 is very basic, it is cell-associated after secretion and is bound avidly by heparin and the extracellular matrix, although it may be released as a soluble form by heparin, heparinase or plasmin. TISSUE SPECIFICITY: The VEGF189, VEGF-165 and VEGF-121 isoforms are widely expressed, whereas the VEGF206 and VEGF-145 are uncommon. SIMILARITY: SwissProt: P15692 ## Belongs to the PDGF/VEGF growth factor family. |
| Stem Cell Type |
|
| Product Usage Statements | |
|---|---|
| Quality Assurance | Routinely evaluated in a 5-day growth assay using human umbilical vein endothelial cells Cell Proliferation Assay: The biological activity of this lot of VEGF was determined by its mitogenic activity on human umbilical vein endothelial cells (HUVEC) with an ED50 of about 1.0 ng/mL. Results may vary depending on the cell line used. VEGF will also stimulate endothelial cells from other species. |
| Usage Statement |
|
| Storage and Shipping Information | |
|---|---|
| Storage Conditions | The lyophilized powder is stable for a few weeks at room temperature, but best stored at -20°C for up to one year from date of receipt. After a quick spin, reconstitute with sterile water or 25mM HEPES and 150mM NaCl, pH 7.0. Reconstituted material can be stored at 2°-8°C for one week, but undiluted aliquots at -20°C are recommended. Avoid repeated freeze/thaw cycles. This product is acid stable. Dilute in either 0.1% TFA or 4mM HCl, containing 25ug BSA per ug VEGF to maintain stability. Reducing reagents (such as dithiothrietol or 2-mercaptoethanol) inhibit VEGF activity. |
| Packaging Information | |
|---|---|
| Material Size | 10 µg |